基于温度的酶注射式葡萄糖生物传感器检测方法研究

针对酶注射式葡萄糖生物传感器在实际使用中因为标定液与被测液的温度不同而引起的测量结果不准确问题,提出一种基于温度的葡萄糖浓度检测方法.首先根据酶促反应动力学建立目前酶注射式葡萄糖生物传感器浓度检测模型,之后利用阿伦尼乌斯公式建立温度与浓度检测动力学模型中未知参数之间的关系,并将此关系代入浓度检测动力学模型中,以建立基于温度的浓度检测新模型.此模型以温度与酶促反应的电流初始斜率为输入值,以被测葡萄糖浓度为输出值,利用此模型提出了以反应混合液的温度和反应初始电流斜率推导被测液浓度的检测方法.利用改进的检测方法进行检测,不仅能够降低温差的影响,提高检测的准确性,还可以省略常规检测中的人工标定,避免人工标定所需的取样探头拆卸步骤,更加有利于在线使用.分别在25.0,30.0和42.0℃下检测1.5 mg/mL和2.5 mg/mL葡萄糖溶液,利用原检测方法与基于温度的检测方法进行检测,结果表明,基于温度的检测方法回收率均在95.0％以上,明显优于原检测方法.     

葡萄糖生物传感器检测方法的研究进展

葡萄糖生物传感器以其灵敏度高、选择性好、反应速度快以及稳定性好等优点吸引了许多研究者的关注。 本文将已发表的一些葡萄糖检测方法分为两类：葡萄糖酶生物传感器检测方法与无酶葡萄糖生物传感器检测方法,简要介绍了这2种检测方法的一些研究进展,并对葡萄糖检测方法的发展前景进行了展望。     

发酵过程葡萄糖在线检测系统的研制

发酵工艺中的高温蒸气灭菌过程使得传统"酶膜"方式葡萄糖生物传感器不能在线使用,导致不能实时控制葡萄糖的补加量,进而影响发酵的质量和产率。为了解决这一问题,本研究构建了一套用于发酵过程的葡萄糖在线检测系统。利用研制的耐高温的"酶液"方式葡萄糖生物传感器解决了"酶膜"方式传感器高温后无法正常工作的问题;利用研制的耐高温的透析取样系统解决了在线取样难题。实验表明:本传感器对葡萄糖的灵敏度达到0.259 nA/(mg/L);检出限为0.7 mg/L。对于500 mg/L葡萄糖浓度测量结果的相对标准偏差为2.02%;在0～1000 mg/L范围内,系统响应信号与葡萄糖浓度呈线性关系;本系统与商品化SBA-40E生物传感分析仪离线测量结果相比,呈现显著的线性相关关系。且线性相关方程的斜率k≈1。利用此检测系统对大肠杆菌发酵过程中的葡萄糖浓度进行了在线测量,并与SBA-40E生物传感分析仪离线测量结果进行了对比,变化趋势基本一致。     

Nafion-二茂铁-双酶修饰的葡萄糖传感器

用二茂铁作为过氧化物酶与玻碳电极的电子传递体,通过牛血清白蛋白-戊二醛交联剂把葡萄糖氧化酶和过氧化物酶固定在Nafion-二茂铁修饰玻碳电极上,制备成葡萄糖传感器。由于工作电位低,电活性物质如抗坏血酸、尿酸等对测定无干扰。该传感器的线性范围为5.0×10～(-4)～2.5×10～(-2)mol/L,响应时间小于30s.     

无酶葡萄糖电化学传感器的研究进展

随着各种新型材料的层出不穷及其在葡萄糖电化学传感器方面应用的发展,无酶葡萄糖电化学传感器的研制成为葡萄糖电化学传感器的另一个研究热点.本文综述了近年来无酶葡萄糖电化学传感器的研究进展,重点介绍了电流型无酶葡萄糖传感器所使用的各种电极材料,总结了最近五年各种新型结构材料在该类传感器研制方面的应用,并对无酶葡萄糖电化学传感器发展方向和趋势进行了展望.     

铂基非酶葡萄糖传感器的研究进展

综述了铂基非酶葡萄糖电化学传感器(包括裸铂电极构建的非酶葡萄糖传感器、掺杂其它金属的铂基非酶葡萄糖传感器和纳米材料修饰的铂基非酶葡萄糖传感器)的研究进展;并对其发展趋势进行了展望(引用文献38篇)。     

无酶葡萄糖传感器

葡萄糖传感器在几十年的发展中取得了重大进展,经历了三代基于酶葡萄糖传感器之后,现已进入第四代无酶葡萄糖传感器的发展阶段.本文从基于酶和无酶两类介绍了不同葡萄糖传感器的测试原理,综述了近年来纳米材料在无酶电化学葡萄糖传感器方面应用的主要研究进展,对不同类别纳米材料的制备方法以及所构建传感器的灵敏度、选择性、检测范围和稳定性等进行了评述,分析了制约无酶葡萄糖传感器商业化应用的主要原因.其中,贵金属纳米材料主要讨论了铂、金和钯;过渡金属纳米材料主要讨论了镍、铜以及其氧化物;双金属纳米材料主要讨论了合金和复合物;碳纳米材料主要讨论了单壁(多壁)碳纳米管和石墨烯.此外,本文也对无酶葡萄糖传感器的发展方向和趋势进行了展望.     

基于聚硫堇和纳米银固定酶的葡萄糖生物传感器

用循环伏安法将电子媒介体硫堇电聚合在玻碳电极表面上,使其表面形成均匀的带负电的聚硫堇膜,通过静电吸附作用吸附表面带正电荷的纳米银溶胶,接着通过静电吸附带负电的葡萄糖氧化酶,最后用聚硫堇包埋电极,从而制得性能优良的葡萄糖氧化酶(GOD)生物传感器。实验发现传感器氧化峰电流与葡萄糖的浓度在1.0×10-8~5.0×10-6mol/L(r=0.9963)范围内呈良好线性关系,检出限为5.0×10-9mol/L(S/N=3)。     

Polyurethane Enzyme Membranes for Chip Biosensors

Abstract

The coverage of electrochemical chip sensors based on silicon technology with a polyurethane enzyme membrane is described. After crosslinking of the surface by polyfunctional isocyanates the enzyme membrane shows good adhesion, complete retention of the enzyme molecules, and low diffusional resistance to both analytes and products. Using thin film noble metal electrodes and ion sensitive field effect transistors, glucose and urea sensors with good long term stability and short response time have been prepared.     

离子结合物在二茂铁介体电流型葡萄糖生物传感器中的应用研究

二茂铁及其衍生物由于氧化还原可逆性好,不溶于水,因此可利用其作介体电流型生物传感器的电子转移介体。但该类传感器的重现性、稳定性差。本文首次提出以含(C_6H_5)_4·(CH_3)_4离子缔合物的碳糊电极为基础电极,以二茂铁(FeCp_2)为电子转移介体制成葡萄糖生物传感器。该传感器重现性、稳定性好,响应迅速,测量准确度高。     

New Immobilized Enzyme Membranes for Tailor-Made Biosensors

Abstract

Commercially available polyamide preactivated membranes for immunodiagnostic use were found suitable for the preparation of immobilized enzyme membranes adaptable to biosensors. Membranes with immobilized glucose oxidese as model enzyme, tested with a microprocessor-based device involving an enzymatic electrode, gave excellent results. The extremely simple and fast procedure allows anyone to prepare such bioactive membranes easily, possibly from his own enzyme preparation and within a few minutes set up a specific probe from commercially available sensors.     

基于柔性衬底的ZnO葡萄糖酶电极制备及特性

通过水热法在长有ZnO籽晶层的柔性聚酰亚胺(PI)衬底上生长了整齐的ZnO纳米棒,ZnO纳米棒的晶体结构和表面形貌通过X射线衍射(XRD)、扫描电子显微镜(SEM)等进行表征.通过静电吸附方式,将葡萄糖氧化酶(GOx)固定在其表面.分别对GOx及修饰前后的ZnO纳米棒进行了紫外-可见光谱表征,发现修饰后存在ZnO的吸收峰和GOx的特征吸收峰,表明GOx固定在ZnO表面.通过对修饰样品进行傅里叶变换红外(FTIR)光谱测试发现了与GOx相关的吸收峰,这进一步表明GOx仍保持生物活性.最后在循环伏安曲线的测试中,这种在柔性衬底上制备的生物酶电极表现出非常灵敏的电流响应,为制备柔性葡萄糖生物传感器奠定了实验基础.     

Macroscopically Oriented Magnetic Core-regularized Nanomaterials for Glucose Biosensors Assisted by Self-sacrificial Label

Magnetic core-regular nanostructures composed of magnetite and regular Prussian blue was prepared by self-sacrificial macro-oriented method. Magnetic graphene oxide (MGO) was vertically oriented on the surface of home-made screen-printing electrode with the help of constant magnetic field (CMF).Then regular nanostructured Prussian blue (RPB) was realized by chemical reaction through an aerosol deposition. Finally, glucose oxidase (GOx) was immobilized by glutaraldehyde cross-linking to fabricate glucose biosensors. The linear range of CMF-RPB/MGO sensor towards glucose was 0.03∼1.35 mM, and the detection limit was 13.4 μM. The CMF-RPB/MGO sensor could apply to analyze glucose in human serum samples.     

Sensitive Biosensors Based on (Dendrimer Encapsulated Pt Nanoparticles)/Enzyme Multilayers

A sensitive enzymed‐based biosensor for glucose has been obtained by introducing dendrimer encapsulated Pt nanoparticles via a layer‐by‐layer assembling method. The free amine groups located on each poly(amidoamine) dendrimer molecule were exploited to covalently attach enzyme to the dendrimer chains using carbodiimide coupling. The resultant enzyme electrodes are shown to have excellent sensitivity (as high as 30.33 μA mM^?1 cm^?2) and a limit of detection (about 0.1 μmol L^?1), depending on metal nanoparticles within dendrimers and the biocompatibility of dendrimers, the linear response range to glucose (from 5 μM to 1.0 mM), a fast response time (within 5 s), and good reproducibility (<8% relative standard deviation between electrodes at low substrate concentration). The sensitivities, and stabilities determined experimentally have demonstrated the potential of dendrimer encapsulated Pt nanoparticles as a novel candidate for enzymatic glucose biosensors.     

Highly Selective and Sensitive Amperometric Biosensing of Glucose at Ruthenium-Dispersed Carbon Paste Enzyme Electrodes

Abstract

The selectivity and sensitivity of glucose measurements at carbon-paste based amperometric biosensors are greatly enhanced through the use of ruthenium-dispersed graphite particles. The improved performance is attributed to the substantial lowering of the overvoltage for the reduction of the hydrogen peroxide product. Hence, cathodic measurements of glucose can be caried out at an optimal potential range (-0.15 to +0.20 V). Contributions from easily oxidizable substances (e.g. acetaminophen, ascorbic and uric acids) are eliminated, without the need for mediators or membrane barriers. The electrocatalytic action of the ruthenium sites results also in a substantially improved sensitivity. A fast flow injection operation is illustrated.     

Glucose sensor using a phospholipid polymer-based enzyme immobilization method

An electroenzymatic glucose sensor based on a simple enzyme immobilization technique was constructed and tested. The glucose sensor measures glucose concentrations as changes of oxygen concentrations induced by enzymatic reactions. The immobilizing procedure was developed with the purpose of producing wearable biosensors for clinical use. Two types of biocompatible polymers, 2-methacryloyloxyethyl phosphorylcholine (MPC) copolymerized with dodecyl methacrylate (PMD) and MPC copolymerized with 2-ethylhexyl methacrylate, were compared as a sensitive membrane of biosensors. The PMD enzyme membrane had a better response time. Linearity, reproducibility, effect of the concentrations of immobilized enzyme and drifts of sensor characteristics in long-term tests were also investigated. The linear characteristics were confirmed with glucose concentration from 0.01 to 2.00 mmol/l, with a coefficient of determination of 0.9999. The average output current for 1 mmol/l and the standard deviation were 0.992 and 0.0283 muA. Significant changes in the sensor's characteristics were not observed for 2 weeks when it was kept in a refrigerator at 4 degrees C. Because of the simple procedure, the enzyme immobilization method is not only useful for wearable devices but also other devices such as micro total analysis systems.     

A Stable Enzyme Biosensor Using Concanavalin a Complex of Glucose Oxidase for Glucose Determination

Abstract

The thermal stability of an insoluble concanavalin A (ConA) complex of glucose oxidase (GOD) was researched. The thermal deactivation rate constants of the complexes were obtained. It was found that the GOD-ConA complexes were less sensitive to thermal inactivation than the native enzyme GOD. By using the complexes, ferrocene-mediated enzyme electrodes were constructed. The results suggested that the GOD-Con A complex electrodes had good thermal stability at room temperature.